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Recognition and processing of branched DNA substrates by Slx1-Slx4 nuclease.

Nucleic Acids Res. 2019; 
Gaur Vineet,Ziajko Weronika,Nirwal Shivlee,Szlachcic Aleksandra,Gapińska Marta,Nowotny Ma
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Bacterial Expression System Synthetic genes for Thielavia terrestris Slx1 (Tt-Slx1) and a fragment of Slx4CCD (Tt-Slx4CCD3) (residues 834–936) were codon-optimized for expression in E. coli and obtained from GenScript®. Get A Quote

摘要

Structure-selective endonucleases cleave branched DNA substrates. Slx1 is unique among structure-selective nucleases because it can cleave all branched DNA structures at multiple sites near the branch point. The mechanism behind this broad range of activity is unknown. The present study structurally and biochemically investigated fungal Slx1 to define a new protein interface that binds the non-cleaved arm of branched DNAs. The DNA arm bound at this new site was positioned at a sharp angle relative to the arm that was modeled to interact with the active site, implying that Slx1 uses DNA bending to localize the branch point as a flexible discontinuity in DNA. DNA binding at the new interface promoted a disorder... More

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